Figures

(A) Unsupervised hierarchical clustering of miRNAseq results from 293T cells treated for 24 hours with vehicle (V), imatinib (IM; 10 μM), doxorubicin (D; 0.5 μg/ml), or a combination of imatinib and doxorubicin (I + D). Ninety-three miRs were clustered into the “D, but not IM + D” category. (B) Top five miRNAs from (A) increased by doxorubicin but not by the imatinib + doxorubicin combination. The P values were calculated as described in Materials and Methods. (C and D) Production of miR-34a and miR-34c in MCF7 (C) or HCT116 (D) cells at 24 hours after the indicated treatments with imatinib and/or doxorubicin relative to vehicle treatment. (E) Immunoblotting of total lysates from the indicated HCT116 cells at 24 hours of the indicated treatments. kd, knockdown; GAPDH, glyceraldehyde-3-phosphate dehydrogenase. (F) Production of miR-34c in cells treated as in (E) relative to vehicle treatment. (G) Schematic of the human miR-34b/c gene locus. E1, upstream exon; EST, expressed sequence tag. (H) Expression of miR34b/c-E1 in cells treated as in (E) relative to vehicle treatment. Relative abundance values are means ± SD of at least three independent experiments. *P < 0.05, **P < 0.01, two-tailed t tests within cell line (C, D, F, and H); ***P < 0.001, two-way analysis of variance (ANOVA) test of interaction, with Bonferroni correction for the two main effect tests on the two cell lines (F).